Abstract
Inversion of the G segment in bacteriophage Mu DNA occurs by a site-specific recombination event and determines the host specificity of Mu phage particles produced. Inversion is mediated by a Mu function (Gin). The gin gene has been placed under control of the inducible λ pL promoter and a synthetic Shine-Dalgarno linker upstream of the initiation codon. The Gin protein content in induced cells is boosted to ˜10% of total protein. Partially purified extracts from overproducing strains promote efficient inversion of the G DNA segment in vitro which is visualized by agarose gel electrophoresis of the substrate DNA after cutting with appropriate restriction endonucleases. The in vitro reaction requires Mg2+, a super-coiled DNA substrate and occurs in the absence of exogenous ATP. Inversion from the G(+) to the G(−) orientation is as efficient as the switch from G(−) to G(+).
Keywords: bacteriophage Mu, site-specific recombination, gin gene product, overproducer, inversion in vitro
Full text
PDF






Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Allet B., Bukhari A. I. Analysis of bacteriophage mu and lambda-mu hybrid DNAs by specific endonucleases. J Mol Biol. 1975 Mar 15;92(4):529–540. doi: 10.1016/0022-2836(75)90307-1. [DOI] [PubMed] [Google Scholar]
- Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chow L. T., Kahmann R., Kamp D. Electron microscopic characterization of DNAs of non-defective deletion mutants of bacteriophage Mu. J Mol Biol. 1977 Jul 15;113(4):591–609. doi: 10.1016/0022-2836(77)90224-8. [DOI] [PubMed] [Google Scholar]
- Daniell E., Abelson J., Kim J. S., Davidson N. Heteroduplex structures of bacteriophage Mu DNA. Virology. 1973 Jan;51(1):237–239. doi: 10.1016/0042-6822(73)90385-1. [DOI] [PubMed] [Google Scholar]
- Frank R., Heikens W., Heisterberg-Moutsis G., Blöcker H. A new general approach for the simultaneous chemical synthesis of large numbers of oligonucleotides: segmental solid supports. Nucleic Acids Res. 1983 Jul 11;11(13):4365–4377. doi: 10.1093/nar/11.13.4365. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gheysen D., Iserentant D., Derom C., Fiers W. Systematic alteration of the nucleotide sequence preceding the translation initiation codon and the effects on bacterial expression of the cloned SV40 small-t antigen gene. Gene. 1982 Jan;17(1):55–63. doi: 10.1016/0378-1119(82)90100-7. [DOI] [PubMed] [Google Scholar]
- Grindley N. D., Lauth M. R., Wells R. G., Wityk R. J., Salvo J. J., Reed R. R. Transposon-mediated site-specific recombination: identification of three binding sites for resolvase at the res sites of gamma delta and Tn3. Cell. 1982 Aug;30(1):19–27. doi: 10.1016/0092-8674(82)90007-1. [DOI] [PubMed] [Google Scholar]
- Grotjahn L., Frank R., Blöcker H. Ultrafast sequencing of oligodeoxyribonucleotides by FAB-mass spectrometry. Nucleic Acids Res. 1982 Aug 11;10(15):4671–4678. doi: 10.1093/nar/10.15.4671. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hsu M. T., Davidson N. Structure of inserted bacteriophage Mu-1 DNA and physical mapping of bacterial genes by Mu-1 DNA insertion. Proc Natl Acad Sci U S A. 1972 Oct;69(10):2823–2827. doi: 10.1073/pnas.69.10.2823. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Iserentant D., Fiers W. Secondary structure of mRNA and efficiency of translation initiation. Gene. 1980 Apr;9(1-2):1–12. doi: 10.1016/0378-1119(80)90163-8. [DOI] [PubMed] [Google Scholar]
- Kahmann R. Methylation regulates the expression of a DNA-modification function encoded by bacteriophage Mu. Cold Spring Harb Symp Quant Biol. 1983;47(Pt 2):639–646. doi: 10.1101/sqb.1983.047.01.075. [DOI] [PubMed] [Google Scholar]
- Kamp D., Chow L. T., Broker T. R., Kwoh D., Zipser D., Kahmann R. Site-specific recombination in phage mu. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):1159–1167. doi: 10.1101/sqb.1979.043.01.131. [DOI] [PubMed] [Google Scholar]
- Kamp D., Kahmann R. The relationship of two invertible segments in bacteriophage Mu and Salmonella typhimurium DNA. Mol Gen Genet. 1981;184(3):564–566. doi: 10.1007/BF00352543. [DOI] [PubMed] [Google Scholar]
- Kamp D., Kahmann R., Zipser D., Broker T. R., Chow L. T. Inversion of the G DNA segment of phage Mu controls phage infectivity. Nature. 1978 Feb 9;271(5645):577–580. doi: 10.1038/271577a0. [DOI] [PubMed] [Google Scholar]
- Krasnow M. A., Cozzarelli N. R. Site-specific relaxation and recombination by the Tn3 resolvase: recognition of the DNA path between oriented res sites. Cell. 1983 Apr;32(4):1313–1324. doi: 10.1016/0092-8674(83)90312-4. [DOI] [PubMed] [Google Scholar]
- Kwoh D. Y., Zipser D. Identification of the gin protein of bacteriophage mu. Virology. 1981 Oct 15;114(1):291–296. doi: 10.1016/0042-6822(81)90280-4. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
- Nash H. A. Integration and excision of bacteriophage lambda: the mechanism of conservation site specific recombination. Annu Rev Genet. 1981;15:143–167. doi: 10.1146/annurev.ge.15.120181.001043. [DOI] [PubMed] [Google Scholar]
- Plasterk R. H., Brinkman A., van de Putte P. DNA inversions in the chromosome of Escherichia coli and in bacteriophage Mu: relationship to other site-specific recombination systems. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5355–5358. doi: 10.1073/pnas.80.17.5355. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Plasterk R. H., Ilmer T. A., Van de Putte P. Site-specific recombination by Gin of bacteriophage Mu: inversions and deletions. Virology. 1983 May;127(1):24–36. doi: 10.1016/0042-6822(83)90367-7. [DOI] [PubMed] [Google Scholar]
- Reed R. R., Grindley N. D. Transposon-mediated site-specific recombination in vitro: DNA cleavage and protein-DNA linkage at the recombination site. Cell. 1981 Sep;25(3):721–728. doi: 10.1016/0092-8674(81)90179-3. [DOI] [PubMed] [Google Scholar]
- Reed R. R. Transposon-mediated site-specific recombination: a defined in vitro system. Cell. 1981 Sep;25(3):713–719. doi: 10.1016/0092-8674(81)90178-1. [DOI] [PubMed] [Google Scholar]
- Remaut E., Stanssens P., Fiers W. Plasmid vectors for high-efficiency expression controlled by the PL promoter of coliphage lambda. Gene. 1981 Oct;15(1):81–93. doi: 10.1016/0378-1119(81)90106-2. [DOI] [PubMed] [Google Scholar]
- Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
- Zieg J., Simon M. Analysis of the nucleotide sequence of an invertible controlling element. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4196–4200. doi: 10.1073/pnas.77.7.4196. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van de Putte P., Cramer S., Giphart-Gassler M. Invertible DNA determines host specificity of bacteriophage mu. Nature. 1980 Jul 17;286(5770):218–222. doi: 10.1038/286218a0. [DOI] [PubMed] [Google Scholar]